PMID- 35066739 OWN - NLM STAT- MEDLINE DCOM- 20221122 LR - 20231007 IS - 1573-6814 (Electronic) IS - 1389-9333 (Print) IS - 1389-9333 (Linking) VI - 23 IP - 4 DP - 2022 Dec TI - Study of the biological characteristics of human umbilical cord mesenchymal stem cells after long-time cryopreservation. PG - 739-752 LID - 10.1007/s10561-021-09973-1 [doi] AB - Human umbilical cord mesenchymal stem cells (hUC-MSCs) have considerable potential in cell therapy. Cryopreservation represents the gold standard in cell storage, but its effect on hUC-MSCs is still not well understood. The aim of this study was to investigate the effect of one year of cryopreservation and thawing on the biological characteristics of hUC-MSCs from the same donors. Fresh hUC-MSCs were cryopreserved in commercial freezing medium (serum-free CellBanker 2) at passage 2. After one year of cryopreservation, the hUC-MSCs were thawed and subcultured to passage 4. The comparison was performed in terms of followings: cell count, viability, morphology, proliferation capacity, differentiation potential and chromosomal stability. The total cell count and viability of hUC-MSCs before and after one year of cryopreservation were 1 x 10(7) and 96.34% and 0.943 x 10(7) and 93.81%, respectively. Cryopreserved and fresh hUC-MSCs displayed a similar cell doubling times, expressed the markers CD73, CD90, CD105 and were negative for the markers CD34, CD45, and HLA-DR. Karyotypes were found to be normal after one year of cryopreservation. The trilineage differentiation properties were maintained after cryopreservation. However, when compared to freshly isolated hUC-MSCs from the same donor, cryopreserved hUC-MSCs exhibited decreased expression of osteogenesis- and chondrogenesis-related genes including Runx2, Sox9, and Col1a1, and increased expression of adipogenesis-related genes. These results demonstrated that cryopreservation did not affect cell morphology, surface marker expression, cell viability, proliferative capacity, or chromosomal stability. However, the osteogenic and chondrogenic differentiation capacities of cryopreserved hUC-MSCs were slightly reduced compared with those of fresh cells from the same donor. CI - (c) 2021. The Author(s). FAU - Zhang, Mingqi AU - Zhang M AD - Department of Stem Cell Center of Precision Medicine Innovation Institute, He University, Hunnan District, No.66 Sishui Street, Shenyang, 110163, China. AD - Liaoning Key Lab of Ophthalmic Stem Cells, He University, Shenyang, China. AD - Liaoning Province Ophthalmic Stem Cell Clinical Application Research Center, He Eye Hospital, Shenyang, China. FAU - Zhao, Yan AU - Zhao Y AD - Department of Stem Cell Center of Precision Medicine Innovation Institute, He University, Hunnan District, No.66 Sishui Street, Shenyang, 110163, China. AD - Liaoning Key Lab of Ophthalmic Stem Cells, He University, Shenyang, China. FAU - Wang, Le AU - Wang L AD - Department of Stem Cell Center of Precision Medicine Innovation Institute, He University, Hunnan District, No.66 Sishui Street, Shenyang, 110163, China. AD - Liaoning Key Lab of Ophthalmic Stem Cells, He University, Shenyang, China. FAU - Zheng, Yuqiang AU - Zheng Y AD - Department of Stem Cell Center of Precision Medicine Innovation Institute, He University, Hunnan District, No.66 Sishui Street, Shenyang, 110163, China. AD - Liaoning Key Lab of Ophthalmic Stem Cells, He University, Shenyang, China. FAU - Yu, Hui AU - Yu H AD - Department of Stem Cell Center of Precision Medicine Innovation Institute, He University, Hunnan District, No.66 Sishui Street, Shenyang, 110163, China. AD - Liaoning Key Lab of Ophthalmic Stem Cells, He University, Shenyang, China. FAU - Dong, Xiaoming AU - Dong X AD - Department of Stem Cell Center of Precision Medicine Innovation Institute, He University, Hunnan District, No.66 Sishui Street, Shenyang, 110163, China. AD - Liaoning Key Lab of Ophthalmic Stem Cells, He University, Shenyang, China. FAU - He, Wei AU - He W AD - Department of Stem Cell Center of Precision Medicine Innovation Institute, He University, Hunnan District, No.66 Sishui Street, Shenyang, 110163, China. AD - Liaoning Key Lab of Ophthalmic Stem Cells, He University, Shenyang, China. AD - Liaoning Province Ophthalmic Stem Cell Clinical Application Research Center, He Eye Hospital, Shenyang, China. FAU - Yin, Zhengqin AU - Yin Z AD - Department of Stem Cell Center of Precision Medicine Innovation Institute, He University, Hunnan District, No.66 Sishui Street, Shenyang, 110163, China. qinzyin@aliyun.com. FAU - Wang, Zhuoshi AU - Wang Z AD - Department of Stem Cell Center of Precision Medicine Innovation Institute, He University, Hunnan District, No.66 Sishui Street, Shenyang, 110163, China. hsyk2017@163.com. AD - Liaoning Key Lab of Ophthalmic Stem Cells, He University, Shenyang, China. hsyk2017@163.com. LA - eng GR - 20-301-4-00/shenyang science and technology bureau/ GR - 2020-BS-292/department of science and technology of liaoning province/ GR - RC190172/shenyang science and technology innovation talents support project/ PT - Journal Article DEP - 20220123 PL - Netherlands TA - Cell Tissue Bank JT - Cell and tissue banking JID - 100965121 SB - IM MH - Humans MH - *Mesenchymal Stem Cells MH - Chondrogenesis MH - Cryopreservation/methods MH - Umbilical Cord MH - Chromosomal Instability PMC - PMC9675661 OTO - NOTNLM OT - Cell proliferation OT - Cryopreservation OT - Human umbilical cord mesenchymal stem cells OT - Karyotype OT - Multilineage differentiation OT - Surface marker COIS- The authors have no conflicting financial interest EDAT- 2022/01/24 06:00 MHDA- 2022/11/23 06:00 PMCR- 2022/01/23 CRDT- 2022/01/23 20:38 PHST- 2021/07/01 00:00 [received] PHST- 2021/10/21 00:00 [accepted] PHST- 2022/01/24 06:00 [pubmed] PHST- 2022/11/23 06:00 [medline] PHST- 2022/01/23 20:38 [entrez] PHST- 2022/01/23 00:00 [pmc-release] AID - 10.1007/s10561-021-09973-1 [pii] AID - 9973 [pii] AID - 10.1007/s10561-021-09973-1 [doi] PST - ppublish SO - Cell Tissue Bank. 2022 Dec;23(4):739-752. doi: 10.1007/s10561-021-09973-1. Epub 2022 Jan 23.